Search Results (210)

The escalating consumption of red meat is a potent environmental risk factor for inflammatory bowel disease (IBD), which is characterized by compromised expression of the xenobiotic transporter P-glycoprotein (MDR1/ABCB1). While gut microbiota metabolize dietary tryptophan into diverse indole derivatives that function as aryl hydrocarbon receptor (AhR) ligands, their differential regulation of MDR1 remains an unresolved AhR paradox. Here, we investigated the mechanisms by which two distinct metabolites, indole-3-acetic acid (IAA) and skatole, regulate MDR1 expression in human colonic epithelial Caco-2 cells. We observed that IAA selectively enhances MDR1 protein stability via an AhR-dependent pathway without inducing de novo transcription, suggesting a mechanism we term enhanced proteostasis mediated by the AhR-Hsp90 complex. Conversely, skatole, a toxic dysbiotic metabolite linked to red meat intake, triggered a time-dependent depletion of MDR1 and potently abrogated the protective efficacy of IAA. Our findings are consistent with a model in which skatole acts as a putative structural disruptor, potentially destabilizing the chaperone complex essential for MDR1 integrity. This destruction is facilitated by a key bacterial enzyme, indoleacetate decarboxylase (IAD), which is a pH-dependent metabolic switch in the gut. The modern Western diet, characterized by high protein and low fiber content, elevates colonic pH, thereby activating IAD to convert protective IAA into toxic skatole. These findings provide a molecular framework for the red meat–microbiome–barrier failure axis and highlight the restoration of the IAA/skatole balance through dietary intervention as a promising therapeutic strategy. Full article

This study aimed to quantify horizontal and vertical bone gain using superimposition of preoperative and postoperative cone beam computed tomography (CBCT) in severe alveolar ridge defects treated with a modified guided bone regeneration (GBR) technique based on customized titanium occlusive barriers with a window design, combined with autologous blood clot and β-tricalcium phosphate (β-TCP). In this prospective case series, 13 patients (28 defects) were treated. Customized titanium barriers were digitally designed based on CBCT data and manufactured by laser sintering. The barriers were fixed over the defects and filled with a mixture of an autologous blood clot and β-TCP, providing an osteoconductive scaffold within a stable regenerative compartment. A standardized window-based follow-up protocol was applied during healing, including irrigation and controlled deepithelialization. Primary outcomes were horizontal and vertical bone gain, assessed by pre- and postoperative CBCT superimposition. Histological evaluation was performed at the time of implant placement. After 8 months, significant bone gain was observed, with a mean horizontal gain of 4.50 ± 2.02 mm and a mean vertical gain of 4.40 ± 2.82 mm (p < 0.0001), confirmed by linear mixed-effects models and patient-level sensitivity analyses (p < 0.001). Histological analysis revealed well-vascularized newly formed bone with active osteoblasts and no inflammatory response. Keratinized gingiva formation was observed at all sites. One minor complication (mild screw loosening) was recorded and successfully resolved. This study is presented as a prospective case series; therefore, the results should be interpreted as exploratory evidence and do not allow direct comparisons or conclusions regarding equivalence or superiority over other GBR techniques. The present report specifically evaluates the regenerative phase prior to functional loading; therefore, although implants were placed according to protocol, implant survival and long-term functional outcomes were not assessed and cannot be inferred from these data. Within the limitations of this prospective case series, customized titanium occlusive barriers with a window design demonstrated promising results for horizontal and vertical bone augmentation and keratinized gingiva formation, without the need for autologous bone grafts or primary wound closure. Full article

Prostate cancer remains largely refractory to immunotherapy, implying the existence of context-specific immune landscape programs that diverge between circulation and tumor. Here, we integrate bulk RNA sequencing from three cohorts (patient peripheral mononuclear cells, primary prostate tissue, and biochemical-recurrence tumors) with multiparameter flow cytometry, unsupervised UMAP/T-REX (Tracking Responders Expanding) mapping, and de novo discovery of long non-coding RNAs (lncRNAs) to characterize context-specific immunoregulation. Patient PBMCs revealed a coherent IL-1/TNF/IL-17 inflammatory architecture with strong chemotactic programs and an unexpected neutrophil-like signal despite density-gradient isolation, consistent with low-density PMN-MDSCs. In contrast, tumors broadly repressed chemokines and innate immune mediators, yet upregulated prostate cancer-associated lncRNAs, indicating local immune quiescence coupled with non-coding regulatory programs. Recurrent tumors acquired epithelial–mesenchymal transition and metabolic remodeling, accompanied by relapse-associated lncRNA signatures, whereas long-term nonrecurrent tumors preserved epithelial and stress-response networks. High-dimensional cytometry confirmed discrete, cancer-enriched myeloid clusters expressing CD47, SIRPα, PD-L1, CD73, and Galectin-9. Network analysis highlighted inflammatory hubs (CXCL2, PTGS2) in PBMCs and loss of mechanotransduction modules in tumors. Structural modeling uncovered a three-way junction and 3′ triple helix in lncRNA. Collectively, these data suggest that circulating inflammatory rewiring is associated with checkpoint-rich suppressor expansion and tumor immune quiescence, outlining integrated myeloid- and RNA-directed strategies for cancer research. Full article

Lineage plasticity, the ability of cancer cells to alter their differentiated state through transcriptional and epigenetic reprogramming, has emerged as a key mechanism of therapeutic resistance across cancers. This adaptive process can manifest in multiple ways, including epithelial–mesenchymal transition, acquisition of stem-like features, and histological transformation, the most striking and clinically apparent example. In EGFR-mutant lung adenocarcinoma (LUAD), lineage plasticity is increasingly recognized as a prevalent mechanism of acquired resistance to tyrosine kinase inhibitors (TKIs). Among its visible manifestations, histologic transformation into small-cell lung cancer (SCLC) is the most frequent, while squamous transformation and other phenotypic shifts also occur. Transformed tumors typically retain the initiating EGFR mutation but lose EGFR dependence, acquire neuroendocrine features, and display aggressive clinical behavior with poor clinical outcomes compared with both de novo SCLC and non-transformed LUAD. Recent studies show that plasticity arises through combined genomic, transcriptomic, and epigenetic reprogramming, often foreshadowed by molecular alterations before overt histological change. Spatial and single-cell profiling reveal heterogeneous trajectories and intermediate states, while functional models and multi-omics approaches have begun to identify therapeutic vulnerabilities distinct from both de novo EGFR-mutated SCLC and classical EGFR-mutated LUAD. Thus, lineage plasticity, whether manifested as histologic transformation or through more subtle epigenetic reprogramming, represents a formidable resistance mechanism in NSCLC. Defining its molecular basis and temporal dynamics will be essential for early detection, prognostication, and the development of tailored therapies. Full article

Background: Colorectal cancer (CRC), particularly the microsatellite-stable (MSS) subtype, remains largely unresponsive to immune checkpoint inhibitors (ICIs) due to immune escape, tumor-associated macrophage (TAM) enrichment, and cytokine-driven suppression that sustain a TAM-dominant tumor microenvironment (TME). To overcome these barriers, a pH-responsive solid lipid nanoparticle (SLN) system was engineered to co-deliver CB-5083 (a VCP/p97 inhibitor), miR-142 (a PD-L1-targeting microRNA), and imiquimod (R, a TLR7 agonist) for spatially confined induction of endoplasmic reticulum stress (ERS) and immune reprogramming in MSS CRC. Methods: The SLNs were coated with PEG–PGA for pH-triggered de-shielding and functionalized with PD-L1- and EGFR-binding peptides plus an ER-homing peptide, enabling tumor-selective and subcellular targeting. Results: The nanoplatform displayed acid-triggered PEG–PGA detachment, selective CRC/TAM uptake, and ER localization. CB-mediated VCP inhibition activated IRE1α/XBP1s/LC3II, PERK/eIF2α/ATF4/CHOP, and JNK/Beclin signaling, driving apoptosis and autophagy, while miR-142 suppressed PD-L1 expression and epithelial–mesenchymal transition markers. R facilitated dendritic cell maturation and M1 polarization. Combined CB + miR + R/SLN-CSW suppressed IL-17, G-CSF, and CXCL1, increased infiltration of CD4⁺ and CD8⁺ T cells, reduced Tregs and M2-TAMs, and inhibited tumor growth in CT-26 bearing mice. The treatment induced immunogenic cell death, reprogramming the TME into a T cell-permissive state and conferring resistance to tumor rechallenge. Biodistribution analysis confirmed tumor-preferential accumulation with minimal off-target exposure, and biosafety profiling demonstrated low systemic toxicity. Conclusions: This TME-responsive nanoplatform therefore integrates ERS induction, checkpoint modulation, and cytokine suppression to overcome immune exclusion in MSS CRC, representing a clinically translatable strategy for chemo-immunotherapy in immune-refractory tumors. Full article

Background/Objectives: Lavandula stoechas has reported bioactivities, but its selective anticancer potential in human models remains insufficiently defined. This study aimed to compare cytotoxicity and selectivity of ethanol and methanol extracts prepared from fresh and dried L. stoechas and to profile candidate bioactive metabolites. Methods: Aerial parts Lavandula stoechas L. subsp. stoechas (L. stoechas L.) were extracted with ethanol or methanol from fresh (LsFE, LsFM) and dried (LsDE, LsDM) material. Cytotoxicity was assessed in cancer (MDA-MB-231, T98G, RT4) and non-malignant (hGF, ARPE-19) cells using Hoechst 33342-stained nuclear counts and MTS viability at 24–48 h. Metabolite identification was performed using LC–QTOF–MS in both positive and negative ESI modes, supported by database search results. Results: All extracts reduced viability in a dose- and time-dependent manner. Among them, the ethanol extract from fresh material (LsFE) displayed the highest cytotoxic potency and the most favorable selectivity profile, markedly reducing viability in breast (MDA-MB-231) and glioblastoma (T98G) cells while exerting only mild effects on non-malignant fibroblast (hGF) and retinal epithelial (ARPE-19) cells. In contrast, extracts from dried material, particularly LsDE, showed broader cytotoxicity across both cancerous and non-cancerous lines. LC–MS highlighted sesquiterpenoids (Kikkanol A; 3(4→5)-Abeo-4,11:4,12-diepoxy-3-eudesmanol), phenolics (tyrosol; 3,4-dihydroxybenzoic acid), flavonoid/ionone derivatives (luteolin 5,3′-dimethyl ether; 3-hydroxy-β-ionone), oxidized fatty acids (9(S)-HpODE, α-EpODE, 5,12-dihydroxy-eicosatetraenoic acid), and jasmonates (12-hydroxyjasmonic acid; dihydrojasmonic acid methyl ester), especially enriched in LsFE. Conclusions: Ethanol extracts of L. stoechas L., especially LsFE, demonstrated selective cytotoxicity against cancer cells while exerting relatively mild effects on non-malignant cells. The metabolite profile of L. stoechas L. extracts revealed a diverse composition, including phenolics, terpenoids, flavonoids, and oxidized lipids, which are commonly associated with biological activity. These results suggest that LsFE is a promising candidate for further studies focusing on compound isolation and mechanistic analysis. Full article

Background: In the bovine mammary gland, de novo fatty acid synthesis is a critical process for milk fat production, in which acetyl-CoA synthetase 2 (ACSS2) serves as a key enzyme by converting acetate into acetyl-CoA. This metabolic pathway is intricately regulated by non-coding RNAs, particularly through the competitive endogenous RNA (ceRNA) mechanism.Purpose: To elucidate the regulatory role and molecular mechanism of the circRNA-02213/miR-328/ACSS2 axis in the lipid metabolism of bovine mammary epithelial cells (BMECs). Methods: Bioinformatic prediction and dual-luciferase reporter assays were employed to verify the targeting interactions among circRNA-02213, miR-328, and ACSS2. In BMECs, qRT-PCR, Western blot, triglyceride/cholesterol quantification, Oil Red O staining, and cell proliferation assays were used to evaluate the effects of this axis on key lipid-metabolic indices and cellular phenotypes. Results: circRNA-02213 functioned as a molecular “sponge” that sequestered miR-328, thereby upregulating ACSS2 expression. Functionally, circRNA-02213 overexpression markedly promoted triglyceride and cholesterol synthesis, lipid droplet accumulation, and BMEC proliferation; whereas miR-328 exerted significant inhibitory effects on these lipid-metabolic processes and cell proliferation. Conclusions: This study demonstrates that circRNA-02213 acts as a ceRNA to relieve miR-328-mediated repression of ACSS2, constituting a critical network that regulates milk fat synthesis and metabolism. The circRNA-02213/miR-328/ACSS2 axis represents a potential molecular target for improving milk lipid quality in ruminants. Full article

Fractional laser photothermolysis, long established in dermatology, enables controlled microthermal injury that stimulates repair without scarring, but its potential in oral tissue regeneration has not been systematically explored. In this study, we conducted the first controlled experimental evaluation of a 1550 nm erbium fiber laser for oral mucosa regeneration. Thirty-two rabbits underwent fractional photothermolysis at energy levels of 70, 100 and 130 kJ, with gingival biopsies collected at 1, 14, 28 and 42 days for histological and immunohistochemical assessment of epithelial repair, stromal remodeling, inflammation and angiogenesis. All energy modes produced microcoagulation columns followed by progressive epithelial thickening, fibroblast proliferation and neoangiogenesis. The 70 kJ mode occasionally led to residual fibrosis, whereas higher energies (100–130 kJ) promoted effective connective tissue remodeling and de novo tissue formation without scarring. Complete epithelial recovery occurred within two weeks, indicating a safe and optimal interval for repeated exposure. Overall, the results demonstrate that 1550 nm fractional photothermolysis is a safe and effective method to induce regenerative responses in oral tissues, establishing a foundation for its translational application in periodontal and peri-implant regeneration. Full article

Background and Objectives: Adequate peri-implant soft tissue dimensions are essential for health, hygiene, and esthetics. When ridge volume is sufficient, phenotype modification may avoid bone grafting. This case report describes a pedicled roll flap performed concurrently with single-stage implant placement after spontaneous socket healing, without bone substitute, and assesses soft-tissue stability with serial intraoral scans. Clinical case: A single-tooth edentulous site underwent prosthetically driven, fully guided implant placement. A modified roll flap with vertical and palatal incisions was prepared; the de-epithelialized crestal connective tissue was elevated and rolled into a buccal envelope to augment thickness. No graft material was used. A provisional crown conditioned the emergence profile. Follow-up included photographs, radiographs, and intraoral scan superimpositions at 2 weeks, 3–4 months, 8 months, and 14 months after implant treatment. Healing was uneventful. Buccal soft-tissue thickness increased, keratinized mucosa was preserved, and midfacial levels remained stable. Emergence profile and papillae integrated harmoniously. Crestal bone levels were stable radiographically. Digital scans corroborated soft-tissue thickness maintenance. No donor-site morbidity occurred. Conclusions: In healed sockets with adequate bone, a modified pedicled roll flap at implant placement can thicken the peri-implant phenotype and achieve stable esthetic integration without bone substitutes. Full article

Aeromonas veronii is a major pathogen threatening freshwater aquaculture, yet the molecular mechanisms of Pelteobagrus vachellii’s immune response to this infection remain unclear. This study integrated histopathology, mRNA-seq and small RNA-seq to investigate P. vachellii’s response to A. veronii at 48 h post-challenge. Histopathologically, infection induced gill epithelial detachment, hepatocyte swelling with cytoplasmic vacuolation, and melanomacrophage centers (MMCs) in the mid-kidney (histological assessment of the head kidney was not feasible due to sampling limitations associated with its small size). Transcriptomic analysis identified 1210 differentially expressed genes (DEGs) in the head kidney (819 downregulated, 391 upregulated), significantly enriched in 11 immune pathways (e.g., NF-κB, Th17 cell differentiation, Complement and coagulation cascades), with key immune genes (e.g., IL-1β, TCRα, CCL4) upregulated. Gene Set Enrichment Analysis (GSEA) revealed activation of the proteasome, ribosome and oxidative phosphorylation pathways, and suppression of the autophagy-animal, FoxO and AMPK pathways. Small RNA-seq identified 544 known and 958 novel miRNAs in the head kidney, with 42 downregulated and 36 upregulated differentially expressed miRNAs (DE miRNAs). The miRNA-mRNA network showed that DE miRNAs (e.g., miR-101-y/z, miR-132-z, miR-3167-y) negatively regulated immune-related target genes (IL-1R1, IRF4, IκBα) in core immune pathways. Collectively, this study clarifies the pathological and miRNA-mRNA regulatory modules of P. vachellii head kidney against A. veronii infection, providing valuable information that enables the further analyses of the defense mechanisms of P. vachellii against A. veronii infection. Full article

Chronic wounds are considered a silent epidemic that impact millions of human lives worldwide, causing comorbidities, reducing life quality and expectancy. Diabetic, pressure, and venous ulcers are the three major clinical entities of chronic wounds, in which the presence of a chronicity phenotype and episodes of recurrence remain as contemporary challenges. We are, accordingly, far from a full understanding about the potential endogenous, predisposing factors that may drive both chronicity and recurrence. Decades of academic and financial endeavors have not translated into a pharmacological intervention that may curb these events. These wounds may exhibit the clinical aspect of a torpid granulative response, poor angiogenesis, delayed or abnormal re-epithelialization, and low contraction rates. At the cellular level, chronicity is propelled and distinguished by the triad of interplaying loops of inflammation, oxidative stress, and cellular senescence. Although the proximal molecular drivers of chronicity and their hierarchal debut sequence are a critical research target and pending task, our unifying hypothesis behind chronicity and recurrence is founded on the existence of an epigenetic pathologic code that originates and perpetuates a “chronic wound memory”. In vitro studies suggest that this de novo edited script is sheltered in dermal fibroblasts and keratinocytes and is spreadable and transmissible to descendant cells, dictating abnormal traits even in ideal culture conditions and successive passages. The list of epigenomic alterations and their significance in wound pathology is continuously escalating. The accurate identification of the key epigenetic priming codes of impaired healing, and their selective re-editing, will be remarkably beneficial. Full article

Background: Atezolizumab plus bevacizumab and lenvatinib are standard treatments for advanced hepatocellular carcinoma, but conventional tumor markers such as alpha-fetoprotein and des-gamma-carboxy prothrombin have a limited ability to reflect treatment responses. Circulating tumor cells with cancer stem cell characteristics have emerged as promising biomarkers. We examined the dynamics of cancer stem cell-related circulating tumor cell subsets and tumor markers at early and maximal response phases in patients with unresectable hepatocellular carcinoma undergoing systemic therapy. Methods: Sixty-two patients treated with either atezolizumab plus bevacizumab or lenvatinib were retrospectively analyzed. Peripheral blood was collected at baseline, during the early phase (during one to three months), and at maximal response. Circulating tumor cell subsets expressing cancer stem cell markers (CD90, epithelial cell adhesion molecule; CD133, vimentin) were assessed using multiparametric flow cytometry and compared with alpha-fetoprotein and des-gamma-carboxy prothrombin. Results: Early decreases in CD90-positive circulating tumor cells after therapy were associated with tumor shrinkage, longer periods of progression-free survival in both groups, and prolonged overall survival in the atezolizumab plus bevacizumab group. By contrast, early changes in alpha-fetoprotein and des-gamma-carboxy prothrombin were not consistently related to tumor size, progression-free survival, or overall survival. At maximal response, changes in CD90-positive circulating tumor cells reflected tumor burden more accurately than alpha-fetoprotein or des-gamma-carboxy prothrombin. Conclusions: These findings indicate that cancer stem cell-related circulating tumor cell subsets, particularly CD90-positive cells, may serve as valuable biomarkers for monitoring treatment response and predicting prognosis in unresectable hepatocellular carcinoma. CD90-positive circulating tumor cells perform dynamic monitoring superior to conventional markers such as alpha-fetoprotein and des-gamma-carboxy prothrombin. Full article

Plant-derived compounds offer immense therapeutic potential, yet many suffer from limited solubility, instability, and poor bioavailability, restricting their clinical application. Curcumin, a polyphenol extracted from Curcuma longa, is one such molecule, with proven antioxidant and anti-inflammatory properties. To overcome its pharmacokinetic limitations, we developed Jamamina, a sustainable nanostructured lipid carrier (NLC) system incorporating curcumin and a Natural Deep Eutectic Solvent (NaDES) phase composed of malic acid and betaine. The bioinspired formulation, based on Amazonian tucumã butter and jambu oil, achieved high encapsulation efficiency (>80%) and curcumin amorphization, enhancing solubility and colloidal stability. In vitro assays with L132 demonstrated potent antioxidant activity (DPPH), a significant reduction in pro-inflammatory cytokines (TNF-α and IL-6), and upregulation of IL-10. The system also suppressed MMP-2/9 activity and preserved cytoskeletal integrity under oxidative stress. These findings highlight Jamamina as a multifunctional, eco-friendly nanoplatform that enables the pharmacological application of plant-derived curcumin, representing a promising platform for modulating redox balance and investigating inflammation in epithelial-like contexts. Full article

Background: Inhaled delivery of cannabidiol (CBD) through dry powder inhalers is a promising approach for achieving optimal drug concentrations in the lungs. Spray drying is a commonly employed technique to prepare inhalable powders with particle sizes ideally ranging from 1 to 5 µm, for deep lung deposition. However, formulating aerosolizable CBD dry powders remains challenging due to the thermolabile nature of CBD and the cohesive behaviour of micron-sized particles, which affects powder dispersibility, reduces de-agglomeration during inhalation, and causes inefficient lung deposition. These challenges can be overcome by the inclusion of excipients that can stabilize CBD during processing and enhance the dispersion and aerosolization of the powder. Objectives and methods: This study investigates the role of different amino acids (lysine, cysteine, arginine, and phenylalanine) in combination with inulin, a sugar-based excipient, on the in vitro aerosolization performance, stability, and cytotoxicity of inhalable CBD dry powders. Results and conclusion: The prepared CBD dry powders exhibited a size range of 1–5 µm. Amino-acid-free CBD powder showed an irregular and flaky morphology, while in association with amino acids, CBD dry powder showed spherical morphology with a dimpled surface. The ATR-FTIR spectra confirmed no interactions between CBD and amino acids in the dry powder formulations. CBD dry powder formulations containing amino acids demonstrated a better aerosolization profile compared to amino-acid-free CBD powder, with the lysine-containing formulation achieving the highest fine particle fraction (FPF) of 56.6%. Additionally, all the formulations were stable under low and high humidity (<15% RH and 53% RH) conditions for 28 days. Cytotoxicity studies on A549 alveolar basal epithelial cells showed that the amino acids were non-toxic, while the CBD formulations with/without amino acids showed comparable levels of cytotoxicity. Full article

Human Papillomavirus (HPV), particularly high-risk strains such as HPV16 and HPV18, is a leading cause of cervical cancer and a significant risk factor for several other epithelial malignancies. While the oncogenic mechanisms of viral proteins E6 and E7 are well characterized, the broader effects of HPV infection on host transcriptional regulation remain less clearly defined. This study explores the hypothesis that conserved genomic motifs within the HPV genome may act as molecular decoys, sequestering human transcription factors (TFs) and thereby disrupting normal gene regulation in host cells. Such interactions could contribute to oncogenesis by altering the transcriptional landscape and promoting malignant transformation.We conducted a computational analysis of the genomes of high-risk HPV types using MEME-ChIP for de novo motif discovery, followed by Tomtom for identifying matching human TFs. Protein–protein interactions among the predicted TFs were examined using STRING, and biological pathway enrichment was performed with Enrichr. The analysis identified conserved viral motifs with the potential to interact with host transcription factors (TFs), notably those from the FOX, HOX, and NFAT families, as well as various zinc finger proteins. Among these, SMARCA1, DUX4, and CDX1 were not previously associated with HPV-driven cell transformation. Pathway enrichment analysis revealed involvement in several key biological processes, including modulation of Wnt signaling pathways, transcriptional misregulation associated with cancer, and chromatin remodeling. These findings highlight the multifaceted strategies by which HPV may influence host cellular functions and contribute to pathogenesis. In this context, the study underscores the power of in silico approaches for elucidating viral–host interactions and reveals promising therapeutic targets in computationally predicted regulatory network changes. Full article